Methanonaphthalene compositions and methods of using the same



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as 3&4009230 w p x 31 Umted Sta METHANONAI'HTHALENE COMPOSITIONS ANDMETHODS OF USING THE SAME Hsing Yun Fan and William E. Rader, Modesto,Calif., Marvin Legator, Rockville, Md., and Linda C. Larrick, Modesto,Calif., assignors to Shell Oil Company, New York, N.Y., a corporation ofDelaware No Drawing. Continuation-impart of abandoned application Ser.No. 87,991, Feb. 9, 1961. This application Nov. 18, 1963, Ser. No.324,223

14 Claims. (Cl. 424-283) ABSTRACT OF THE DISCLOSURE Methods andcompositions for controlling microorganisms and molluscs by contactingthem with a 2,3,4a,8adiepoxy-polyhydronaphthalene-5,8-dione such as6,7-dichloro exo 2,3 endo 4a,8a diepoxy1,2,3,4,4a,8ahexahydro-exo-l,4-methanonaphthalene-5,8-dione.

This application is a continuation-in-part of copending application,Ser. No. 87,991, filed Feb. 9, 1961, now abandoned.

This invention pertains to the control of mjgoorganisms and molluscswith polycyclic diepoxides, morefi= ticularly described aspolyhydronaphthalene diones containing two epoxide rings, one of theseepoxide rings involving the carbon atoms in the 4a and 8a positions, andthe other involving the carbon atoms in the 2 and 3 positions. Thesecompounds have the essential character represented by the structuralFormulae I and II.

In terms of name, these compounds can be designated as2,3,4a,Sa-diepoxypolyhydronaphthalene-S,S-diones. Included in thisgeneral class of diepoxides are those wherein a methano bridge joins thecarbon atoms at the 1- and 4-positions. These, of course, are the2,3,4a,8a-diepoxypolyhdro-1,4-methanonaphthalene-5,8-diones.

Described more particularly, the compounds contemplated by thisinvention :are characterized by the structural formula:

XII I XI o X-i-X 0| XII X l wherein each X, X and X" is individuallyselected from the group consisting of hydrogen, middle halogen (i.e.,bromine and chlorine) and low molecular weight organic groups, with theproviso that at least one of X" is hydrogen. The broken lines in theformula indicate the fact that this class of novel compounds includesthose having a methano bridge between the carbon atoms in the number 1-and number 4-positions, as well as those having no such bridge, andthose compounds having an ethylenic or double bond between the carbonatoms in the number 6 and 7 positions, as well as those which have asaturated or single bond between those same two carbon atoms.

The organic groups represented by the symbols X, X' and X",respectively, preferably are low molecular weight 1 Sept. 3, 1968hydrocarbon or substituted hydrocarboni.e., groups containing not morethan about 10 carbon atoms each-and the compounds of greatest interestare those wherein each of the organic groups represented by thesesymbols is a hydrocarbon group. Such hydrocarbon groups may be of eitheraliphatic or cyclic configuration; they may be saturated, olefinicallyunsaturated or aromatically unsaturated; preferably they are free fromacetylenic unsaturation. The aliphatic groups may be of straight-chainor of branched-chain configuration. The aromatic groups preferably aremononuclear. Thus, suitable organic groups include both straight-chainand branched-chain alkyl such as methyl, ethyl, nand isopropyl, n, sec-,and tert-butyl, the various isomeric C C and like alkyl groups,cycloalkyl such as the cyclopentyl, cyclohexyl, methylcyclohexyl,dimethylcyclooctyl, 3,5,5-trimethylcyclohexyl, and like cycloalkylgroup's, aryl such as the phen= yl group, alkaryl such as themethylphenyl, ethylphenyl and like alkaryl groups, aralkyl such as thebenzyl, phenethyl and like aralkyl groups, alkenyl such as the allyl,crotyl and like groups, alkadienyl such as the butadienyl, pentadienyland like alkadienyl groups, and mixed groups such as the vinylphenyl,allylphenyl, phenylvinyl, phenylcrotyl, phenylallyl groups, and thelike. Of the substituted hydrocarbon groups, those hydrocarbon groupsset out above which are substituted by one or more of halogen, cyano andnitro are preferred. Because of their properties, the compounds whereinthe organic groups represented by the symbols X, X, and X are alkyl offrom 1 to 6 carbon atoms are preferred.

Of these compounds, the subclass wherein there is a 1,4-methano bridge,an olefinic double bond between the carbon atoms in the 6- and7-positions, and at least one of X is middle halogen are of particularinterest because of their high toxicity toward microorganisms. In termsof name, these preferred compounds can be designated as2,3,4a,8a-diepoxy-polyhydro-1,4-methanonaphthalene-5,8-diones6,7-olefinically unsaturated and containing halogen on at least one ofthe carbon atoms at the 6 and 7 positions. From the available data, itappears that the 6,7-di(middle halo) compounds of this subclass have thehighest toxicity toward a broad spectrum of microorganisms.

Since starting materials for the preparation of such compounds whereinall of X and X are hydrogen are at present most readily available andthe resulting compounds exhibit high activity toward microorganisms, thecompounds of this class wherein all of X and X are hydrogen arepreferable. These compounds would, of course, be the1,2,3,4,4a,8a-hexahydro members of the class.

The compounds contemplated by this invention can exist in the form ofstereoisomers, of two general types: one in which the two epoxy ringsare in cis configuration, and one in which the two epoxy rings are intrans configuration. That is, if the carbon atoms of the hexane ,ringare considered to lie in the same plane, both of the epoxy rings lie onthe same side of that plane, or the two epoxy rings can lie on oppositesides of that plane, respectively. (As used herein, the termstereoisomer designates only the geometric isomers whose spatialconfiguration differs, and does not include optical isomers isomer pairswhich exist because one is the mirror image of the other.)

In the compounds of this class in which no methano bridge is present,there are thus two series of stereoisomers. However, in the compounds inwhich a methano bridge is present, there are four series ofstereoisomers. This results because, as is shown in US. Patent No.2,717,851, in a bicycloheptane structure, the methano bridge does notlie in the same plane as the carbon atoms of the hexane ring. Thus, inthe compounds of this class wherein a methano bridge is present eaeh ofthe epoxy 3 rings can be of cis or trans configuration with respect tothe methano bridge, and also can be of cis or trans configuration withrespect to the other epoxy ring. In terms of more conventionalnomenclature employed with bridged. rings, these compounds can beexo-2,3:exo-4a,8adiepoxy, or exo-2,3:endo-4a,8a-diepoxy, orendo-2,3:exo- 4a,8a-diepoxy, or endo-2,3:endo-4a,8a-diepoxy inconfiguration.

Typical species of the compounds include:

6,7-dichloro-exo-2,3 :-endo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-exo-1,4-methanonaphthalene-5,8-dione;

-6,7dichloro-exo-2-,3 :exo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-endo-l,4-methanonaphthalene-5,8-dione;

6,7-dichloro-endo-2,3 :ex-4a,8a-diepoxy-1,2,3,4,4a8a,--hexahydro-endo-1,4-methanonaphthalene-5,8-dione;

6,7 -dichloro-endo-2,3 :end0-4a,8a-die-poxy- 1 ,2,3,4,4a,8ahexahydro-exo-l,4-methanonaphthalene-S,8-dione;

Cis- 2,3 :4a,8a-diepoxy)decalin-5 ,8-dione;

Trans- 2,3 :4a,8a-diepoxy decalin-S ,8-dione;

6,7 -dichloro-cis(2,3 :4a,8a-diepoxy) -1,2,3,4,4a,6,7,8a-

octahydronaphthalene-S ,8-dione;

6,7-dibromo-trans- (2,3 :4a,8a-diepoxy)-1,2,3,4,4a,6,7,8a=-

octahydronaphthalene-5,-8dione;

7-methyl-cis (2,3 4a,8a-diepoxy)decalin-S, 8-dione;

Z-methyl-trans- (2,3 :4a,8a-diepoxy)decalin-5,8-dione;

Exo-2,3 :exo-4a,8a-diepoxy-1,2,3,4,4a,8a-hexahydroendo-l,4-methanonaphthalene-5,S-dione;

Exo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,6,7,8a-octahydro-exo-1,4-methanonaphthalene-5,8-dione;

7-methyl-exo-2,3 :endo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-exo-1,4-rnethanonaphthalene-5,8-dione;

Z-methyl-exo-Z, 3 :exo-4a,8a-diepoxy- 1 ,2,3,4,4a,8a-hexahydro-endo-l,4-methanonaphthalene-5,8-dion'e;

2,6,7-trichloro-endo-2,31exo-4a,8a-diepoxy-1,2,3,4,4a,8ahexahydro-endo-l,4-methanonaphthalene-5,8-dione;

1,2,4,6,7,9,9-heptachloro-end-2,3 :endo-4a,8a-diepoxy-1,2,3,4,4a,8a-hexahydro-exo-1,4-methanonaphthalene- 5,8-dione;

1,4,7,9,9-pentabromo-exo-2,3:exo-4a,8a-diepoxy-1,2,3,4,4a,8a-hexahydro-endo-1,4-methanonaphthalene-5,8- dione;

2,6-dimethyl-exo-2,3 :endo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-exo-1,4-methanonaphthalene-5,8-dione;

1,4,9,9-tetrachloro-endo-2,3 :exo-4a,8a-diepoxy-1,2,3,4(4a,8a-hexahydro-endo-1,4-methanonaphthalene-5,8- dione;

2,7-dibromo-exo-2,3 :exo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-endo-1,4-methanonaphthalene-5,8,-dione;

2,6-dichloro-exo-2,3 :exo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-endo-l,4-methanonaphthalene-S,8-dione;

1,4,6,7-tetrabromo-endo-2,3 :endo-4a,8a-diepoxy-1,2,3,4,4a,8a-hexahydro-exo-1,4-methanonaphthalene-5,8- dione;

I,2,4,6,9,9-hexachloro-endo-2,3 :exo-4a,8a-diepoxy-1,2,3,4,4a,8a-hexahydro-endo-1,4-methanonaphthalene-5,8- dione;

7-methyl-exo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,6,7,8aoctahydro-exo-l,4-methanonaphthalene-5,8-dione;

6-phenyl-exo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,6,7,8aoctahydro-exo-1,4-methanonaphthalene-5,8-di0ne;

2-benzyl-endo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,6,7,8aoctahydro-exo-1,4-methanonaphthalene-5,8-dione;6-chloro-exo-2,3 :endo-4a,8a-diepoxy-l,2,3,4,4a,8a-

hexahydro-exo-1,4-methanonaphthalene-5,8-dione; 6-bromo-endo-2,3:exo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-endo-l,4-methanonaphthalene-5,8-dione; 6-methyl-exo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,8a-

hexahydro-exo-1,4-methanonaphthalene-5,8-dione; 6-chloro-exo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,6,7,8a-

octahydro-exo-1,4-methanonaphthalene-5,8-dione;6-chloro-7-methyl-exo-2,3 :endo-4a,8a-diepoxy-1,2,3,4,4a,

8a-hexahydro-exo-1,4-methanonaphthalene-5,8-dione.

Preferred methods for preparing the compounds con-- templated by thisinvention involve epoxidation of the corresponding di-ethylenicallyunsaturated compoundsthat is, the corresponding polyhydronaphthalene5,8- diones and the corresponding1,4-dihydro-1,4-methanonaphthalene-5,8-diones. Copending applicationSer. No. 162,965 filed Dec. 26, 1961, now Patent No. 3,225,070,discloses 4a,8a' epoxy 4a,8a dihydronaphthalene-5,8- diones, and methodsfor their preparation. These 4a,8aepoxides can be directly epoxidized tothe corresponding 2,3:4a,8a-diepoxy compounds of this invention. Fordetails of the preparation of the 4a,8a-epoxy precursors, reference ishereby made to said US. Patent No. 3,225,- 070. Briefly, however, these4a,8a-epoxy precursors can be prepared as follows:

Step I.A suitable butadiene or cyclopentadiene compound is reacted in aDiels-Alder type of synthesis with a suitable halogenated quinone.

Step II.The product of Step I is reacted with acetic acid and zinc, andthen with chromic acid.

These two steps are described in detail in the Journal of the AmericanChemical Society, vol. 76, pages 6150 (1954).

Step III.The product of Step II is epoxidized by reaction with hydrogenperoxide in the presence of sodium carbonate or other water-soluble,alkaline material at about room temperature.

Conduct of this epoxidation is set out in detail in US. Patent No.3,225,070. As is shown therein, in those compounds containing a methanobridge, the product can be in either of two stereoisomeric formsone inwhich the methano bridge and epoxy ring are cis, and one in which themethano bridge and epoxy ring are trans, with respect to the bridgering.

This method results in the compounds of this invention wherein thecarbon atoms in the 6 and 7 positions are joined by an olefinic doublebond. Compounds wherein the carbon atoms in the 6 and 7 positions aresaturated can be prepared by hydrogenation of the olefinic double bond.Orthodox methods for effecting hydrogenation of olefinic double bondscan be used to effect the hydrogenation in this case also.

Step IV.The diepoxides are prepared from the monoepoxides by means ofacid epoxidation, employing a peracid as the epoxidizing agent. Theconduct of the epoxidation is wholly in accord with the well knownprinciples for conducting the peracid epoxidation of an isolatedolefinic double bond, these principles being discussed in the article bySwern, Chemical Reviews, vol. 45, pages 1- 68 (1949). The mostconvenient peracids are performic acid, peracetic acid, perbenzoic acidand monoperphthalic acid.

In general, about one mole of the peracid is used per mole of themonoepoxide, although in some cases it may be desirable to employ asmall excesssay, up to about 1020 percent excessof the peracid.Preferably the epoxidation is carried out in a suitable mutual solventfor the reactants and product. Chloroform is an especially usefulsolvent for the purpose, but other materials such as diethyl ether,dichloromethane, benzene, ethyl acetate, etc., and the like, may beused. The temperature employed for effecting the epoxidation may varyover a consider able range, depending upon the particular reactantsused. Generally, suitable temperatures will be found to be within therange of from about 20 C. to about C. Often ordinary roomtemperature-that is, from about 10 C. to about 30 C.will be foundsuitable. In some cases, it will be found desirable to mix the reactantsat a lower temperature, then heat the mixture somewhat to effectcompletion of the reaction. The diepoxide product obtained by thismethod can be recovered from the crude reaction mixture by anyconvenient means known to the art, such as distillation, extraction,fractional precipitation or the like.

The evidence indicates that the major products of this process are thediepoxides having the two epoxide rings on opposite sides of the commoncyclohexane ring-that is, the compounds containing no methano bridgehaving the trans-diepoxy configuration, and the compounds containing themethano bridge having the exo 2,3:endo4a,8a-diepoxy-exo-1,4-methaneconfiguration.

Steps III and IV can be inverted. In this case, the major products arethe diepoxides wherein the two epoxide rings lie on the same sides ofthe common cyclohexane ringthe products being those with the cis-diepoxyconfiguration, including those with theendo-2,3:endo-4a,8a-diepoxy-exo-1,4-methario configuration.

To further illustrate these methods for preparing the compounds of thisinvention, and the appropriate materials and conditions to be used, thefollowing exam= ples show the preparation of a typical member of themost preferred compounds that is, those compounds wherein there is anolefinic double bond between the carbon atoms in the 6- and 7 positions,and each of those carbon atoms is bonded to a middle halogen atom.

Example I A solution of 65 grams of a mixture of isomers of 6,7-dichloro4a,8a epoxy 1,4,4a,8a-tetrahydro 1,4- methanonaphthalene-5,8-dione,prepared according to the method set forth in U.S. Patent No. 3,225,070,in 390 milliliters of chloroform was stirred into a solution of 3 gramsof sodium acetate in 65 milliliters of 40% peroxyacetic acid and themixtur was allowed to stand for 23 hours at room temperature. Themixture then was diluted with water and the separated water layer wasextracted with chloroform. The chloroform solution was dried overanhydrous sodium sulfate and evaporated almost to dryness by means ofair blowing. The resulting residue was triturated with methanol to give55 grams of product which was mainly 6,7-dichloro-exo-2,3:endo-4a,8a-diepoxy-1,2,3,4,4a,8a-hexahydro exo 1,4-methanonaphthalene-5,8-dione.Pure material was obtained by recrystallization from chloroform,chromatography over alumina in methylene chloride, and successiverecrystallizations from ethyl acetate and benzene; M.P. 195- 196 C.

Analysis for C C O C1 percent.Calculated: C, 48.3; H, 2.2; CI, 26.0.Found: C, 48.5; H, 2.6; CI, 26.0.

A second crop (9.5 grams) from the first recrystallization wastriturated with chloroform to leave 0.8 gram of a solid which wasidentified as 6,7-dichloro-endo-2, 3:endo 4a,8a-diepoxy 1,2,3,4,4a,8ahexahydro-exo- 1,4-methanonaphthalene-5,8-dione by comparison of itsinfrared spectrum with that of the product of Example II.

In another experiment the single isomer, 6,7-dichloroendo-4a,8a-epoxy1,4,4a,8a tetrahydro-exo 1,4 methanonaphthalene-S,S-dione, M.P. 118-119C., gave upon epoxidation with peroxyacetic acid a mixture whichinfrared analysis indicated to contain about 80% of the above diepoxideand 20% of the diepoxide of Example II. MMR data showed that themonoepoxide of M.P. 118-1l9 C., the major component of the mixture ofisomers used in the first experiment, has its epoxide group orientedendo. On the basis of the known course of exo-epoxide formation inbicycloheptene systems, it is our belief that the 2,3-epoxy group of thediepoxide of M.P. 195196 C. is oriented exo. Since the diepoxide ofExample II also formed from the monoepoxide of MP. l18-119 C., itsdifference apparently lies in the orientation of the 2,3-epoxy groupwhich is then endo. It will be recognized that just as the monoepoxideof M.P. 118-l19 C. is capable of yielding two diepoxides, in a similarway its stereoisomeric monoepoxide is capable of yielding two otherdiepoxides. These four stereoisomeric diepoxides are indicated to bepresent in the crude product of the first experiment described above.

Example II 21.3 milliliters of tritluoracetic anhydride was addedportionwise to an ice-chilled suspension of 35 milliliters of 90%hydrogen peroxide in 25 milliliters of methylene dichloride over a-minute period, with stirring. The

mixture then was stirred at the temperature of melting ice for another15 minutes, and then was added dropwise with stirring to a mixture of24.1 grams of 6,7 dichloro 1,4 dihydro 1,4 methanonaphthalene 5,8- dioneand 42.5 grams of dry sodium carbonate in milliliters of methylenedichloride, over a period of 40 minutes. After the addition wascomplete, the resulting mixture was heated to reflux and refluxed for 35minutes. The inorganic salt was removed by decantation. The liquid wasevaporated to near dryness and the residue was then triturated withmethanol to give 13.9 grams of yellow solid, melting point 157-161" C.This material was recrystallized twice from ethyl acetate and once frombenzene to give 4.3 grams of 6,l-dich1oro-endo-2,3-epoxy- 1,2,3,4tetrahydro 1,4 methanonaphthalene 5,8- dione, melting at 182-183 C.

Analysis for C H O Cl percent.Calculated: C, 51.3; H, 2.3; Cl, 27.6Found: C, 51.6; H, 2.7; Cl. 27.7.

A solution of 2.14 grams of sodium carbonate in one milliliter and 25.7milliliters of 30% hydrogen peroxide solution in water was added inportions over a 5-minute period with vigorous stirring at roomtemperature to a suspension of 25 .7 grams of the epoxide in 257milliliters of 1,2-dimethoxyethane. The reaction temperature rose to 50C. and then slowly returned to room temperature. After 4 hours, thereaction mixture was diluted with water to precipitate 16.7 grams ofproduct which was mainly 6,7 dichloro-endo 2,3:endo 4a,8a-diepoxy1,2,3,4, 4a,8a hexahydro exo 1,4 methanonaphthalene 5,8- dione.Successive recrystallizations from ethyl acetate and benzene affordedpure material melting at 176-177" C.

Analysis for C C O Cl percent.Calculated: C, 48.3; H, 2.2; CI, 26.0.Found: C, 47.9; H, 2.6; Cl, 26.2.

The designation of the isomeric structure of this diepoxide is discussedin Example I. Since the 2,3-epoxy group of this product is believed tobe endo, then the corresponding precursor mono-epoxide of M.P. 182-183C. described above may be designated as having an endo-2,3-epoxyconfiguration.

The diepoxides of the present invention are highly toxic with respect tomicroorganisms, including fungi, bacteria, and molds. These compoundsare active in controlling pathogenic microorganisms, the causativeagents of disease in warm-blooded animals, such as livestock anddomestic animals, as well as controlling microorganisms attacking plantsand plant materials. Since these diepoxides are substantially nontoxicto plants, including their seeds, when used at the dosages effectiveagainst microorganisms present on those plants, these diepoxides can beused as foliage fungicides and as seed-treating agents.

To demonstrate the effectiveness of these diepoxides as foliagefungicides, a typical species of these diepoxides- 6,7 dichloro exo 2,3endo 4a,8a diepoxy 1,2,3,4, 4a,8a hexahydro exo 1,4 methanonaphthalene5,8- dione, M.P. -196" C.-was tested against eleven foliage pathogens onvarious plant hosts. The pathogens and hosts used were as follows:

Pathogen Host Phytopzthom infestans (light blight)- Potato (SOZIIMMILtuberosum). Phutophthora phaseoli (downy mil- Lima bean (Phaseolushmatus) Stemphylium sultmi (gray leaf spot)- Tomato.

Helminthnsporium turcicum (blight) Corn (7ea maus).

Colletofrichum lagenarium (anthrae- Cucumber (Cucumus sativus).

nose

.Erysiphe cichoracearum (powdery mildew).

Cucumber.

The test was carried out according to the following procedure. The testchemicals were prepared as 1% w./v. stock solutions in a solvent. Thestock was diluted for spraying, using a diluent consisting of distilledwater plus solvent (1:1) or diluent consisting of distilled water plustwo co-solvents (2:1:1). Triton X-155, at 0.0050.1% w., was used as a'wetting agent for each concentration of toxicant. All chemicals wereinitially tested at a single concentration of 1000 ppm. Disease controlbeing shown, the chemicals were retested at 1000 ppm, and the nextlowest dilution of 500 ppm. The chemicals were then retested in thisfashion at successively lower concentrations until the minimum effectiveconcentration was determined.

With the mist-type spray and the diluent mixture which was applied, adrying interval of one to two hours was sufficient before inoculationwith spore suspensions using an atomizer. Inoculations with bean mildewwere made by dusting conidia over treated plants in the greenhouse.

After 24 hours incubation at 19 C. and 100% humidity, the plants wereremoved to the controlled environment greenhouse for symptomdevelopment. Disease control and phytotoxicity assessments were madewithin seven days. Of the greatest significance is the fact thatcompounds of the present invention are outstanding fungicides yet do notmanifest significant phytotoxicity.

The following table summarizes the results of screening:

TABLE I Minimum effective concentration of test compound (parts per 1Test Compound A6,7-diehloro-exo-2,3:endoiafia-diepoxy-l,2,3,4.4aSa-hexahydro-exo-l,4-methanonaphthalone-5, 8-d10ne, melting at 195-196 Clest Compound B6,7-dichloro-endo-2,3zendo;4a,8a-diepoxy-13,3,4,4atEhexahydro-exo-1,4-methanonaphthalene-5,8-dione, melting at 176- 177In no case was there observed phytotoxicity of the test compound towardthe host plants.

Further tests were made to determine the persistence of test compound AvApplication was made with a sprayer to field grown potato plants. At24-hour intervals shoots were cut from these plants, placed in a 5percent sucrose solution and inoculated with spores of Phytophthora infestans. The following table gives the time in days for the testcompound to lose activity. Varying concentrations of the test compoundwere used, as shown in the table.

TABLE II Concentration of Time in days for test compound, test compoundlb./100 gallons: to lose activity 1/ 2 5 It is accordingly evident thatthe test compound is quite persistent, providing protection for from 4to 5 days after its application.

The effectiveness of compounds of this invention against typicalsoil-inhabiting fungi which attack both dormant and germinating seedswas shown by the results of the fol lowing tests: Test compound A wasapplied to sugar beet seeds at two dosages and the seeds planted in asoil containing damping-off fungi consisting of Pythium and Rhizoctoniaspecies. 10-14 days after planting, the seeds were examined to determinethe percent germinated and percent destroyed by the fungii.e.,damped-off. Checks & were also made of untreated soil and of soilsterilized by steaming. The results:

TABLE III Percent germination at dosage 01- Similar results wereobtained on the treatment of cotton seed.

For horticultural purposes, the active compounds of the presentinvention may be used alone or in combination with other fungicidal,viricidal, insecticidal or acaricidal materials, the action on which maybe either internal or external, with plant nutritives, plant hormones,and the like. Wetting agents and, if necessary or desirable, stickerssuch as the heavy hydrocarbon oils with a minimum viscosity of 10 Englerat 50 C. can be present. The wetting agent must be non-reactive with thecompounds of the present invention. Non-ionic surfactants seempreferable. If the toxic agents are employed inthe form of emulsions orsuspensions, for example, in water, solvents such as oils, emulsifiers,emulsion stabilizers, and the like may be added. Materials whichsuppress phytotoxic action may also be added if desired. For example,glucose is known to protect tomato plants against damage by certainsubstances having a phytotoxic effect when employed in concentratedform. The compounds of the present invention may be applied by means ofspraying. Spraying of the plants to be treated may be performed withaqueous emulsions, solutions, or suspensions of the active agents. Thespray liquid is generally applied at a rate of from about to 150 gallonsper acre. If spraying is effected with smaller quantities of liquid asin low-volume spraying, high concentrations of the active agents shouldbe employed. If desired, a minor amount of the order of about 0.001 toabout 0.5% by weight of a wetting agent may be added to aid in forming asuspension in the aqueous medium. Any of the conventional wetting agentscan be employed. Particularly suitable wetting agents are the sodiumsalts of a mixture of secondary heptadecyl sulfate, sold commerciallyunder the names of Teepol and polyethylene glycol ethers of alkylphenols sold under the trade names of Triton X- and Triton X155."Preferably concentrated compositions comprising an active compound ofthe present invention and a suitable wetting agent are prepared, and theconcentrate is then dispersed in water prior to use. 1

A further form in which the fungicidal compounds of the presentinvention may be applied for horticultural use consists of solutions ofthe active ingredient in suitable inert liquid or semi-solid diluents inwhich the active ingredient is present in molecularly dispersed form.The form in which the agents to be employed are applied to the objectstreated depends on the nature of the object and the purpose of theapplication. Suitable inert solvents for the manufacture of liquidpreparations should not be readily flammable, as odorless as possibleand without any toxic effect on humans and animals when properly used.Neither should they have a corrosive effect on the components of thepreparations or the material of the storage vessel. Examples of suitablesolvents are high" boiling oils, e.g., oils of vegetable origin, such ascastor oil, etc., and lower boiling solvents with a flash point of atleast 30 C., such as carbon tetrachloride, ethylene dichloride,acetylene tetrachloride, hydrogenated naphthalene, alkylatednaphthalene, sorbent naphtha, etc. Mix tures of solvents may also beused. Non-aromatic petro leum oils and xylene are commonly employed.

For horticultural use the active compounds of the pres ent invention mayalso be applied in the form of dusts, utilizing as the inert vehiclesuch materials as tricalcium phosphate, precipitated chalk, bentonite,kaolin, kieselguhr, etc. These compounds may also be employed. in theform of aerosols. For this purpose the active ingredient is dissolved ordispersed in a solvent boiling below room temperature at atmosphericpressure.

Diepoxides of the invention are effective slimicides, as is shown by thefact that at a concentration of 25 parts per million by weight6,7-dichloro-exo-2,3:endo-4a,8adiepoxy 1,2,3,4,4a,8a hexahydro exo 1,4methano-= naphthalene-5,8-dione destroyed slime-forming organisms in anindustrial white water obtained from an operating paper mill. The testswere performed by placing a sample of the white water and sufficient ofthe test compound to give the desired concentration, incubating themixture at 37 C. for 48 hours, then streaking Emersons agar with some ofthe incubated mixture, incubating the streaked medium for 48 hours at 37C., then observing the medium to determine the presence or absence ofslime growth.

The diepoxides are employed as slimicides in the manner and by thetechniques known to practitioners of the art. In general, it isnecessary only to add one or more of the diepoxides to provide theslimicidally effective dosage in the water to be treated. In some cases,this dosage will be as little as 5 parts per million parts by weight ofthe water, while in other cases a dosage of as much as 100 parts, or upto 250 parts, of the diepoxide, on the same basis, may be required.

These diepoxides also have been found to be effective molluscicides, lowconcentrations in water killing aquatic molluscs such as snails livingtherein. Thus, it has been found that at a concentration of 3 parts permillion by weight of the water, 6,7-dichloro-exo-2,3:endo-4a,8adiepoxy1,2,3,4,4a,8a hexahydro exo 1,4 methanonaphthalene-5,8-dione gaveessentially complete kill of Helisoma trivolvis living therein, at anexposure time of two hours, and at a concentration of 2 parts .permillion on the same basis, and an exposure time of 24 hours, gavecomplete kill of the snails.

These diepoxides are employed as molluscicides by disseminating them inthe required concentration in the water in which the undesired molluscsare dwelling. Any suitable means for effecting the dissemination can beusedfor example, the diepoxides can be stirred into the water, injectedin a portion of the water wherein the water is in turbulent flow, orlike mechanical means can be used. The dissemination can also beeffected through the use of a highly hydrophilic surface-active agent,such as the water-soluble non-ionic surface-active agents, watersolubleanionic surface-active agents, particularly the esters of sulfuric acid,and which contain a plurality of ether moieties, and the like.

The nonionic surface-active agents which can be used are described inSchwartz and Perry, Surface Active Agents (Interscience, 1949) inchapter 8 thereof, and in Schwartz, Perry and Berch, Surface ActiveAgents (Interscience, 1958), on pages 125-138 and 163-166.

Thus, as pointed out in the first of these references, the suitablesurface-active agents are those water-soluble materials which containether linkages, ester linkages or amide linkages, or which containcombinations of these linkages, to the solubilizing groups. Best knownare the reaction products of hydrophilic hydroxy compounds (phenols,alcohols, including certain glycols) with several moles of a loweralkylene oxide (usually ethylene and/or propylene oxide). The kinds ofthese materials which are known, and methods for their preparation aredescribed in detail in these references. Of particular importance arethe polyethenoxy compounds which are described on pages 125-138 of thesecond reference. Also important are the alkylene oxide block copolymersdescribed on pages 163-166 of that reference.

The diepoxide-surface-active agent compositions can be introduced intothe water to be treated in any convenient manner. Often, particularlywhere the body of water to be treated is small or is in the form of asmall canal, the composition to be used can be introduced by means of asyringe, or hand-pump, or the like. Techniques of introduction whichcause the composition to be introduced in the form of small dro pletsasby, for example, the use of spray nozzles-are preferred.

At least about one part of the diepoxide per milli n parts by Weight ofthe water treated generally is required to effect control of aquaticsnails within a reasonable time, and ordiarily not more than 1000 partsper million of the diepoxide will be required. A concentration of fromabout two to about one hundred parts per million of the diepoxideordinarily will be found satisfactory. At such concentrations, residencetimes of from a few minutes--say, 5-10 minutes-41p to several hoursasmuch as a dayusually are sufficient, with the shorter times beingassociated with higher diepoxide concentrations.

The diepoxides are effective killers of molluscs, and particularlywater-dwelling snails, including Taphius glabratus, Helisomla trivolvis,Lymnrtea bulimoides, Marisa cornuarietis, Pomacea lineara, P. glauca,and Ocinebm japonica.

In addition to being excellent agents for destroying various pests, thecompounds of this invention also are useful for production of polymersof desirable characteristics. For example, useful hard insoluble polymerresins are prepared by heating one or more of the compounds of theinvention at a temperature of about with an equivalent amount of an acidanhydride in the presence of a tertiary amine a-s curing agent.

The compounds of the present invention are active against a broadspectrum of microorganisms and bacteria in addition to the foliage andsoil-inhabiting fungi already discussed. As a consequence of this uniqueactivity, the compounds of the present invention may be employed toprevent or control infection in animals. The following in vitro testresults are illustrative of the usefulness of compounds of thisinvention when used for such purposes.

Test Compound A-6,7-dichlor0-ext2,3:endo-ta,8a-diepoxy-1,2,3,4, ggtwhexahydro-exo-l,4-methanonaphthalene-5,8-dione, melting at 3 TestCompound B-6,7-dichloro-endo--2,3:endo-4a, 8a-diepoxy-1,2,3, 4. ea, sahxahydro-exo-i,i-methanonaphthalene-5,8-dione, melting at It is evidentthat the test compound was effective against all of these bacteria andfungi. Of particular interest is the inhibitory action of this compoundagainst P. aeraginosa, one of the hardest bacteria to attack chemically.

Compounds of this invention have been shown to be active in vivo tests,When mice were injected with compound A. at intervals prior to challengewith a highly infective strain of Staphylococcus aureus, a high percentof the animals were afforded protection from the organism. The followingin vivo test results are illustrative of the usefulness of the compoundsof this invention when used for such purposes.

amazon TABLE v Number of mice surviving out of 10 when compound A lprecedes infection of Staphylococcus aureus hy Dose, lug/kg.

Test Compound A6,7-dichloro-ex-2,3zendo taxa-diepoxy-1,2,3,4, 4a,gihcxahydro-exo-l,t-methanonaphthalene-5,8-drune, inciting at 195- 196Thus compounds of this invention are of value in the prophylaxis andtreatment of certain diseases of warmblooded animals, such as livestockand domestic animals. According to the present invention compositionssuitable for treatment of animals include in addition to the activecompounds of the invention pharmaceutical or veterinary carriers whichmay either be solid or liquid in materials. Preparations for oralinjection can be liquids or solids or any combination of these forms,such as syrups, elixirs, emulsions, powders, capsules or tablets.Preparations for administration of the active agent in unit dose cantake the form of compressed powders on tablets or of a powder enclosedin a suitable capsule of absorbable material such as gelatin. Thecompressed powders, tablets, or capsules may also comprise suitableexcipients and/or diluents such as starch, lactose, stearic acid,magnesium stearate, dextrin, polyvinyl pyrrolidine or other diluentsknown to the pharmaceutical formulating art.

Preparations for parenteral administration may be in the form of sterilesolutions or suspensions in such liquids as water, physiological saline,benzyl alcohol, ethyl oleate, corn oil, peanut oil, Carbowax 600 (watersoluble poly-- ethylene, approximate molecular weight 600), Car-bowax400 (water soluble polyethylene, approximate molecular weight 400),glycerol monoleate, Methosil (water solu ble methylcellulose) and thelike, dimethyl sulfoxide, and may contain soluble or insoluble diluentsand/or solid or liquid exeipients.

Compositions according to this invention may be employed as feedsupplements. Compounds of the present invention may be mixed with aninert carrier or diluent. An inert carrier is one that is nonreactivewith respect to the compounds of this invention and may be administeredwith safety to the animals to be treated. The carrier may be one of theingredients of the animal feed or may be attapulgus clay, pyrophylite,bentonite and th like or may be of vegetable origin; corn cob meal,walnut shell meal, blood meal, fish meal and the like. the like or maybe of animal origin; bone meal, oyster shell meal, blood meal, fish mealand the like.

Compounds of this invention may be employed neat, with suitablecarriers, or in combination with other medicants, vitamins, hormones, orgrowth stimulants.

The compounds of this invention may be mixed into the feed of animals,given in their drinking water, administered parenterally or intubatedorally. As little as 0.1 milli' gram per kilogram of body Weight may besufiicient to protect the animal or to control certain pathogenic organisms or as much as 100 milligrams per kilogram of body weight may beemployed. As feed or water additives a dosage of as low as 0.5 part permillion or as high as 500 parts per million of the compound may berequired,

Compositions of compounds of this invention and suitable inert carriers,vehicles, or diluents may contain as little as 0.001% of the activeingredient or as much as 100% of the active compound.

We claim as our invention:

it 2 1. A horticultural composition which comprises a com-- pound of theformula:

wherein each X, X and X" is individually a member of the groupconsisting of hydrogen, middle halogen, hydrocarbon of from one to tencarbon atoms, free from acetylenic unsaturation, and substitutedhydrocarbon from one to ten carbon atoms, free from acetylenicunsaturation, wherein each substituent is a member of the groupconsisting of halogen, nitro and cyano, with the provision that at leastone of X" is hydrogen, and an inert horticultural adjuvant therefor.

2. The composition of claim 1 wherein said compound is 6,7dichloro-exo-2,3-endo-4a,8a-diepoxy1,2,3,4,4a,8ahexahydro-exo-l,4,-methanonaphthalene-5,8-dione.

3. A method of controlling water-dwelling snails which comprisesapplying to the habitat of said snails an efiective quantity of thecompound of the formula:

wherein each X, X and X" is individually a member of the groupconsisting of hydrogen, middle halogen, hydrocarbon of from one to tencarbon atoms free from acetylenic unsaturation, and substitutedhydrocarbon from one to ten carbon atoms, free from acetylenicunsaturation, wherein each substituent is a member of the groupconsisting of halogen, nitro and cyano, with the provision that at leastone of X is hydrogen.

4. The method of claim 3 wherein said compound is 6,7 dichloroexo-2,3-endo-4a,8a-diepoxy-1,2,3,4,4a,8ahexahydro-exo-l,4-methanonaphthalene-5,8-dione.

5. A method of controlling fungi and bacteria in warmblooded animalscomprising introducing into said animals an effective dosage of acompound having the formula:

wherein each X, X, and X is individually a member of the groupconsisting of hydrogen, middle halogen, hydrocarbon of. from one to tencarbon atoms, free from acetylenic unsaturation, and substitutedhydrocarbon from one to ten carbon atoms, free from acetylenic unsaturation, wherein each substituent is a member of the group consisting ofhalogen, nitro and cyano, with the provision that at least one of X" ishydrogen.

6. The method of claim 5 wherein said compound is administered in thefeed of said animals.

7. A method of controlling bacteria and fungi comprising applying to thelocus of said bacteria and fungi an effective quantity of a compoundhaving the formula:

I ll o\ [0 XII (IV) X wherein each X, X, and X" is individually a memberof the group consisting of hydrogen, middle halogen, hydrocarbon of fromone to ten carbon atoms, free from acet ylenic unsaturation, andsubstituted hydrocarbon of from one to ten carbon atoms, free fromacetylenic unsaturation, wherein each substituent is a member of thegroup consisting of halogen, nitro and cyano, with the provision that atleast one X is hydrogen.

8. The method of claim 7 wherein the compound is represented by FormulaI.

9. The method of claim 7 wherein the compound is represented by FormulaII.

10. The method of claim 7 wherein the compound is represented by FormulaIV.

11. The method of claim 7 wherein the compound is represented by FormulaIII.

12. The method of claim 11 wherein X is chlorine in the compoundrepresented by Formula III.

13. The method of claim 12 wherein X and X are both hydrogen in thecompound represented by Formula III.

14. The method of claim 13 wherein the compound represented by FormulaIII is 6,7-dichloro-exo-2,3-endo 4a,8adiepoxy-1,2,3,4,4a,8a-hexahydro-exo-1,4-methanonaphthalene-5,8-dione.

References Cited UNITED STATES PATENTS 12/1960 Gerolt 167-42 12/1961Phillips 16732 X

